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Fixation with PFA requires a denaturing step, see Abreview by M.Schrader.

Target

Function

Forms a channel through the mitochondrial outer membrane and also the plasma membrane. The channel at the outer mitochondrial membrane allows diffusion of small hydrophilic molecules; in the plasma membrane it is involved in cell volume regulation and apoptosis. It adopts an open conformation at low or zero membrane potential and a closed conformation at potentials above 30-40 mV. The open state has a weak anion selectivity whereas the closed state is cation-selective. May participate in the formation of the permeability transition pore complex (PTPC) responsible for the release of mitochondrial products that triggers apoptosis.

Tissue specificity

Heart, liver and skeletal muscle.

Sequence similarities

Belongs to the eukaryotic mitochondrial porin family.

Domain

Consists mainly of a membrane-spanning beta-barrel formed by 19 beta-strands. The helical N-terminus folds back into the pore opening and plays a role in voltage-gated channel activity.

Images

IHC image of ab34726 staining in human liver formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab34726, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

ab34726 at a 1/100 dilution staining VDAC1/Porin in COS7 cells by Immunocytochemistry/ Immunofluorescence.The antibody does not work with a standard fixation for ICC/IF as it does not recognize the non-denatured protein. However, applying a fixation for hidden antigens (modified from Peränen J, Rikkonen M, Kääriäinen L. A method for exposing hidden antigenic sites in paraformaldehyde-fixed cultured cells, applied to initially unreactive antibodies. J Histochem Cytochem. 1993 Mar41(3):447-54) results in very good recognition of VDAC/porin. Denaturing is achieved by Guanidine hydrochloride in 50 mM Tris-HCL pH 7.5 after fixation (3% pFA, 0.05% GA) and permeabilization (0.2% TX-100). The secondary used was a TRITC conjugated donkey anti-rabbit used at a 1/100 dilution.

I have issued a free of charge replacement with a different lot number, but it looks like that this vial was already on the pick list for another order. Therefore, the order is waiting to receive a vial we al...

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement of another batchwith the order numbe...

Thank you for your reply. For broad reactivity, I would recommend our ab34726. This antibody is known to react with all forms of VDACs including VDAC1, VDAC2 (4 isoforms) and VDAC3(2 isoforms). I hope this helps, please let me know if you need any addi...

Thank you for your enquiry. Currently, this antibody (ab34726) is out of stock. However, the good news is that we have some alternative products against the same target protein (VDAC/Porin). I do not know in which species/application your customer...

Thank you for your inquiry.
VDAC1 is a mitochondrial protein and I believe its level of expression is pretty ubiquitous in all tissues, cancerous and normal.
According to the Human Protein Atlas, VDAC1 is expressed in most normal and cancer tis...

Thank you for your inquiry.
Cells should be permeabilized with Triton X-100 and then incubated in 6 N Guanidine HCl in 50mM Tris-HCl, pH 7.5 for 10 minutes at room temperature.
I hope this information is helpful to you. Please do not hesitate ...